Versatile nitrilases: Nitrile-hydrolysing enzymes

Kobayashi, Michihiko; Shimizu, Sakayu

doi:10.1111/j.1574-6968.1994.tb07036.x

Cited by 147 publications

(38 citation statements)

References 43 publications

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“…Nitrilases with different properties that hydrolyze aromatic and aliphatic substrates have been found in Rhodococcus, Alcaligenes, Acinetobacter, Pseudomonas, Acidovorax etc. [5][6][7][8]. The extensive investigations for the selective transformation of nitriles by biocatalysts over the past decade has revealed an impressive number of enzymes, which possess the desired broad substrate spectrum by retaining their exquisite natural specificities with respect to regioand stereo-selectivity.…”

Section: Introductionmentioning

confidence: 99%

Optimization of nitrilase production from Alcaligenes faecalis MTCC 10757 (IICT-A3): effect of inducers on substrate specificity

Nageshwar

Sheelu

Shambhu

et al. 2010

Bioprocess Biosyst Eng

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Microbial nitrilases are biocatalysts of interest and the enzyme produced using various inducers exhibits altered substrate specificity, which is of great interest in bioprocess development. The aim of the present study is to investigate the nitrilase-producing Alcaligenes faecalis MTCC 10757 (IICT-A3) for its ability to transform various nitriles in the presence of different inducers after optimization of various parameters for maximum enzyme production and activity. The production of A. faecalis MTCC 10757 (IICT-A3) nitrilase was optimum with glucose (1.0%), acrylonitrile (0.1%) at pH 7.0. The nitrilase activity of A. faecalis MTCC 10757 (IICT-A3) was optimum at 35 °C, pH 8.0 and the enzyme was stable up to 6 h at 50 °C. The nitrilase enzyme produced using different inducers was investigated for substrate specificity. The enzyme hydrolyzed aliphatic, heterocyclic and aromatic nitriles with different substitutions. Acrylonitrile was the most preferred substrate (~40 U) as well as inducer. Benzonitrile was hydrolyzed with almost twofold higher relative activity than acrylonitrile when it was used as an inducer. The versatile nitrilase-producing A. faecalis MTCC 10757 (IICT-A3) exhibits efficient conversion of both aliphatic and aromatic nitriles. The aromatic nitriles, which show not much or no affinity towards nitrilase from A. faecalis, are hydrolyzed effectively with this nitrilase-producing organism. Studies are in progress to exploit this organism for synthesis of industrially important compounds.

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Section: Introductionmentioning

confidence: 99%

Optimization of nitrilase production from Alcaligenes faecalis MTCC 10757 (IICT-A3): effect of inducers on substrate specificity

Nageshwar

Sheelu

Shambhu

et al. 2010

Bioprocess Biosyst Eng

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“…Products of the reaction are a carboxylic acid and ammonia. Nitrilases have been characterized from microorganisms (for a review, see Kobayashi and Shimizu 1994) and higher plants (Thimann and Mahadevan 1964a, b). In some plants, nitrilases have been implied in the synthesis of the growth hormone, indole-3-acetic acid (IAA) from indole-3-acetonitrile (IAN) (Thimann and Mahadevan 1964a; Bartling et al 1992Bartling et al , 1994Schmidt et al 1996).…”

Section: Introductionmentioning

confidence: 99%

Enzymatic characterization of the recombinant Arabidopsis thaliana nitrilase subfamily encoded by the NIT 2/ NIT 1/ NIT 3-gene cluster

Vorwerk

Biernacki

Hillebrand

et al. 2001

Planta

154

168

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Three of the nitrilase isoenzymes of Arabidopsis thaliana (L.) Heynh. are located on chromosome III in tandem and these genes (NIT2/NIT1/NIT3 in the 5'-->3' direction) encode highly similar polypeptides. Copy DNAs encompassing the entire coding sequences for all three nitrilases were expressed in Escherichia coli as fusion proteins containing a C-terminal hexahistidine extension. All three nitrilases were obtained as enzymatically active proteins, and their characteristics were determined, including a detailed comparative analysis of their substrate preferences. All three nitrilases converted indole-3-acetonitrile (IAN) to indole-3-acetic acid (IAA), albeit, compared to the most effective substrates found, phenylpropionitrile (PPN), allylcyanide, (phenylthio)acetonitrile and (methylthio)acetonitrile, with low affinity and velocity. The preferred substrates are either naturally occurring substrates, which may originate from glucosinolate breakdown, or they are close relatives of these. Thus, a major function of NIT1, NIT2 and NIT3 is assigned to be the conversion to carboxylic acids of nitriles from glucosinolate turnover or degradation. While all nitrilases exhibit a similar pH optimum around neutral, and NIT1 and NIT3 exhibit a similar temperature optimum around 30 degrees C independent of the substrate analyzed (IAN, PPN), NIT2 showed a remarkably different temperature optimum for IAN (15 degrees C) and PPN (35-40 degrees C). A potential role for NIT2 in breaking seed dormancy in A. thaliana by low temperatures (stratification), however, was ruled out, although NIT2 was the predominantly expressed nitrilase isoform in developing embryos and in germinating seeds, as judged from an analysis of beta-glucuronidase reporter gene expression under the control of the promoters of the four isogenes. It is possible that NIT2 is involved in supplying IAA during seed development rather than during stratification.

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“…It has been widely explored for its potential to synthesize industrially important acids from nitriles (Godfredsen et al, 1985 ;Nishise et al, 1987 ;Yamamoto et al, 1990 ;Bhalla et al, 1992 ;Kobayashi and Shimizu, 1994 ;Wu and Li, 2003 ).…”

Section: Nitrile Metabolism In Microbesmentioning

confidence: 98%